Microorganisms for plant pathogen inhibition

ABSTRACT

The invention relates to treatment of a plant with one or more microorganisms for inhibiting a plant pathogen. More particularly, the invention relates to isolated  Bacillus  strains, and strains having all of the identifying characteristics of these strains, and combinations thereof, for a use comprising the above-mentioned use.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority under 35 U.S.C. § 119(e) to U.S.Provisional Application Ser. No. 62/738,208 filed on Sep. 28, 2018, theentire disclosure of which is incorporated herein by reference.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has been filedelectronically in ASCII format and is hereby incorporated by referencein its entirety. Said ASCII copy, created on Dec. 20, 2019, is named60421-299804_SL.txt and is 738 bytes in size.

FIELD OF THE DISCLOSURE

The invention relates to treatment of a plant with one or moremicroorganisms for inhibiting a plant pathogen. More particularly, theinvention relates to isolated Bacillus strains, and strains having allof the identifying characteristics of these strains, and combinationsthereof, for a use comprising the above-mentioned use.

BACKGROUND AND SUMMARY OF THE INVENTION

The present invention relates to microorganisms, such as isolatedBacillus strains for use in inhibiting plant pathogens. Fungal pathogenscause a variety of diseases in plants, including, but not limited to,basal stem rot disease caused by Ganoderma boninense, bud rot caused byPhytophthora palmivora, and wilt caused by Fusarium oxysporum. Thesediseases detrimentally affect palm plants on oil palm plantations, andcause significant economic losses in the palm oil industry. Anadditional fungal species, Cladosporium cladosporioides causesCladosporium rot in the leaves and fruit of many plants, most notablyred wine grapevines. C. cladosporioides affects up to 50% of grapeclusters at harvest, greatly reducing yield and detrimentally impactingwine quality. Yeasts such as Pichia manshurica, which grow on the fruitsurface can contribute to off-odors and wine spoilage also resulting inloss of wine quality and financial loss. Aspergillus fumigatus is aubiquitous mold that is commonly found on decaying vegetation and is notonly a problem as an agent of soft rot, but has also been established asthe global leading cause of aspergillosis in humans and causes one ofthe highest numbers of deaths among patients with fungal infections.Many other fungal diseases have detrimental effects on plants and canuse significant economic losses in the horticulture, agricultural, andforestry industries in general, for example. Ralstonia solanacearum is asoil-borne bacterial strain that causes bacterial wilt in a wide rangeof host plants.

Applicants have developed Bacillus strains, and combinations thereof,that are useful for inhibiting plant pathogens. These strains compriseBacillus strains selected from the group consisting of Bacillus strain86 (NRRL No. B-50944), a strain having all of the identifyingcharacteristics of Bacillus strain 86 (NRRL No. B-50944), Bacillusstrain 300 (NRRL No. B-50943), a strain having all of the identifyingcharacteristics of Bacillus strain 300 (NRRL No. B-50943), Bacillusstrain 101 (NRRL No. B-67218), a strain having all of the identifyingcharacteristics of Bacillus strain 101 (NRRL No. B-67218), Bacillusstrain 235 (NRRL No. B-67219), a strain having all of the identifyingcharacteristics of Bacillus strain 235 (NRRL No. B-67219), Bacillusstrain 77 (NRRL No. B-67274), a strain having all of the identifyingcharacteristics of Bacillus strain 77 (NRRL No. B-67274), Bacillusstrain 177 (NRRL No. B-67275), a strain having all of the identifyingcharacteristics of Bacillus strain 177 (NRRL No. B-67275), Bacillusstrain 102 (NRRL No. B-67276), a strain having all of the identifyingcharacteristics of Bacillus strain 102 (NRRL No. B-67276), Bacillusstrain ATC2 (NRRL No. B-67134), a strain having all of the identifyingcharacteristics of Bacillus strain ATC2 (NRRL No. B-67134), Bacillusstrain A12 (NRRL No. B-67516), a strain having all of the identifyingcharacteristics of Bacillus strain A12 (NRRL No. B-67516), Bacillusstrain V17 (NRRL No. B-67664), a strain having all of the identifyingcharacteristics of Bacillus strain V17 (NRRL No. B-67664), Bacillusstrain V18 (NRRL No. B-67665), a strain having all of the identifyingcharacteristics of Bacillus strain V18 (NRRL No. B-67665), Bacillusstrain 1607 (NRRL No. B-67666), a strain having all of the identifyingcharacteristics of Bacillus strain 1607 (NRRL No. B-67666), andcombinations thereof.

In one embodiment, a method of treating a plant to inhibit a fungal or abacterial disease is provided. The method comprises contacting the plantwith a composition comprising an effective amount of an isolatedBacillus strain selected from the group consisting of Bacillus strain 86(NRRL No. B-50944), a strain having all of the identifyingcharacteristics of Bacillus strain 86 (NRRL No. B-50944), Bacillusstrain 300 (NRRL No. B-50943), a strain having all of the identifyingcharacteristics of Bacillus strain 300 (NRRL No. B-50943), Bacillusstrain 101 (NRRL No. B-67218), a strain having all of the identifyingcharacteristics of Bacillus strain 101 (NRRL No. B-67218), Bacillusstrain 235 (NRRL No. B-67219), a strain having all of the identifyingcharacteristics of Bacillus strain 235 (NRRL No. B-67219), Bacillusstrain 77 (NRRL No. B-67274), a strain having all of the identifyingcharacteristics of Bacillus strain 77 (NRRL No. B-67274), Bacillusstrain 177 (NRRL No. B-67275), a strain having all of the identifyingcharacteristics of Bacillus strain 177 (NRRL No. B-67275), Bacillusstrain 102 (NRRL No. B-67276), a strain having all of the identifyingcharacteristics of Bacillus strain 102 (NRRL No. B-67276), Bacillusstrain ATC2 (NRRL No. B-67134), a strain having all of the identifyingcharacteristics of Bacillus strain ATC2 (NRRL No. B-67134), Bacillusstrain A12 (NRRL No. B-67516), a strain having all of the identifyingcharacteristics of Bacillus strain A12 (NRRL No. B-67516), Bacillusstrain V17 (NRRL No. B-67664), a strain having all of the identifyingcharacteristics of Bacillus strain V17 (NRRL No. B-67664), Bacillusstrain V18 (NRRL No. B-67665), a strain having all of the identifyingcharacteristics of Bacillus strain V18 (NRRL No. B-67665), Bacillusstrain 1607 (NRRL No. B-67666), a strain having all of the identifyingcharacteristics of Bacillus strain 1607 (NRRL No. B-67666), andcombinations thereof, and inhibiting the fungal disease or the bacterialdisease.

In another embodiment, a commercial package is provided. The commercialpackage comprises an isolated Bacillus strain A12 (NRRL No. B-67516), astrain having all of the identifying characteristics of Bacillus strainA12 (NRRL No. B-67516), Bacillus strain V17 (NRRL No. B-67664), a strainhaving all of the identifying characteristics of Bacillus strain V17(NRRL No. B-67664), Bacillus strain V18 (NRRL No. B-67665), a strainhaving all of the identifying characteristics of Bacillus strain V18(NRRL No. B-67665), Bacillus strain 1607 (NRRL No. B-67666), a strainhaving all of the identifying characteristics of Bacillus strain 1607(NRRL No. B-67666), and combinations thereof.

In yet another embodiment, a composition is provided comprising anisolated Bacillus strain selected from the group consisting of Bacillusstrain A12 (NRRL No. B-67516), a strain having all of the identifyingcharacteristics of Bacillus strain A12 (NRRL No. B-67516), Bacillusstrain V17 (NRRL No. B-67664), a strain having all of the identifyingcharacteristics of Bacillus strain V17 (NRRL No. B-67664), Bacillusstrain V18 (NRRL No. B-67665), a strain having all of the identifyingcharacteristics of Bacillus strain V18 (NRRL No. B-67665), Bacillusstrain 1607 (NRRL No. B-67666), a strain having all of the identifyingcharacteristics of Bacillus strain 1607 (NRRL No. B-67666), andcombinations thereof.

The following clauses, and combinations thereof, provide variousadditional illustrative aspects of the invention described herein. Thevarious embodiments described in any other section of this patentapplication, including the section titled “DETAILED DESCRIPTION OFILLUSTRATIVE EMBODIMENTS” and the EXAMPLES are applicable to any of thefollowing embodiments of the invention described in the numbered clausesbelow.

1. A method of treating a plant to inhibit a fungal disease or abacterial disease, the method comprising contacting the plant with acomposition comprising an effective amount of an isolated Bacillusstrain selected from the group consisting of Bacillus strain 86 (NRRLNo. B-50944), a strain having all of the identifying characteristics ofBacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No.B-50943), a strain having all of the identifying characteristics ofBacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No.B-67218), a strain having all of the identifying characteristics ofBacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No.B-67219), a strain having all of the identifying characteristics ofBacillus strain 235 (NRRL No. B-67219), Bacillus strain 77 (NRRL No.B-67274), a strain having all of the identifying characteristics ofBacillus strain 77 (NRRL No. B-67274), Bacillus strain 177 (NRRL No.B-67275), a strain having all of the identifying characteristics ofBacillus strain 177 (NRRL No. B-67275), Bacillus strain 102 (NRRL No.B-67276), a strain having all of the identifying characteristics ofBacillus strain 102 (NRRL No. B-67276), Bacillus strain ATC2 (NRRL No.B-67134), a strain having all of the identifying characteristics ofBacillus strain ATC2 (NRRL No. B-67134), Bacillus strain A12 (NRRL No.B-67516), a strain having all of the identifying characteristics ofBacillus strain A12 (NRRL No. B-67516), Bacillus strain V17 (NRRL No.B-67664), a strain having all of the identifying characteristics ofBacillus strain V17 (NRRL No. B-67664), Bacillus strain V18 (NRRL No.B-67665), a strain having all of the identifying characteristics ofBacillus strain V18 (NRRL No. B-67665), Bacillus strain 1607 (NRRL No.B-67666), a strain having all of the identifying characteristics ofBacillus strain 1607 (NRRL No. B-67666), and combinations thereof, andinhibiting the fungal disease or the bacterial disease.

2. The method of clause 1 wherein the Bacillus strain is selected fromthe group consisting of Bacillus strain 300 (NRRL No. B-50943), a strainhaving all of the identifying characteristics of Bacillus strain 300(NRRL No. B-50943), Bacillus strain 101 (NRRL No. B-67218), a strainhaving all of the identifying characteristics of Bacillus strain 101(NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), a strainhaving all of the identifying characteristics of Bacillus strain 235(NRRL No. B-67219), Bacillus strain 77 (NRRL No. B-67274), a strainhaving all of the identifying characteristics of Bacillus strain 77(NRRL No. B-67274), Bacillus strain A12 (NRRL No. B-67516), a strainhaving all of the identifying characteristics of Bacillus strain A12(NRRL No. B-67516), Bacillus strain 1607 (NRRL No. B-67666), a strainhaving all of the identifying characteristics of Bacillus strain 1607(NRRL No. B-67666), and combinations thereof.

3. The method of clause 1 or 2 wherein the Bacillus strain is selectedfrom the group consisting of Bacillus strain 300 (NRRL No. B-50943), astrain having all of the identifying characteristics of Bacillus strain300 (NRRL No. B-50943), Bacillus strain 235 (NRRL No. B-67219), a strainhaving all of the identifying characteristics of Bacillus strain 235(NRRL No. B-67219), Bacillus strain 1607 (NRRL No. B-67666), a strainhaving all of the identifying characteristics of Bacillus strain 1607(NRRL No. B-67666), and combinations thereof.

4. The method of clause 2 wherein the Bacillus strain is Bacillus strain300 (NRRL No. B-50943).

5. The method of clause 2 where the Bacillus strain is Bacillus strain235 (NRRL No. B-67219).

6. The method of clause 2 wherein the Bacillus strain is Bacillus strain1607 (NRRL No. B-67666).

7. The method of clause 2 wherein the Bacillus strain is Bacillus strain101 (NRRL No. B-67218).

8. The method of clause 2 wherein the Bacillus strain is Bacillus strain77 (NRRL No. B-67274).

9. The method of any one of clauses 1 to 8 wherein the Bacillus strainhas antifungal activity.

10. The method of any one of clauses 1 to 9 wherein the fungal diseaseis caused by a fungus of a genus selected from the group consisting ofGanoderma, Phytophthora, Fusarium, and combinations thereof.

11. The method of clause 10 wherein the fungal disease is caused by afungus selected from the group consisting of Ganoderma boninense,Ganoderma mirabile, Ganoderma weberianum, Phytophthora palmivora,Fusarium kyushuense, Fusarium oxysporum, Fusarium nelsonii, Cladosporiumcladosporioides, Pichia manshurica, Pichia kudriavzevii, Aspergillusfumigatus, and combinations thereof.

12. The method of clause 11 wherein the fungus is Ganoderma boninense.

13. The method of clause 11 wherein the fungus is Ganoderma mirabile.

14. The method of clause 11 wherein the fungus is Ganoderma weberianum.

15. The method of clause 11 wherein the fungus is Phytophthorapalmivora.

16. The method of clause 11 wherein the fungus is Fusarium kyushuense.

17. The method of clause 11 wherein the fungus is Fusarium oxysporum.

18. The method of clause 11 wherein the fungus is Fusarium nelsonii.

19. The method of any one of clauses 1 to 18 further comprising treatingthe plant with a different Bacillus strain, a lactic acid bacterialstrain, and combinations thereof.

20. The method of any one of clauses 1 to 19 wherein the effectiveamount of the Bacillus strain is about 1.0×10² CFU/ml or gram of thecomposition used to contact the plant to about 1.0×10⁸ CFU/ml or gram ofthe composition used to contact the plant.

21. The method of any one of clauses 1 to 19 wherein the effectiveamount of the Bacillus strain is about 1.0×10² CFU/ml or gram of thecomposition used to contact the plant to about 1.0×10⁴ CFU/ml or gram ofthe composition used to contact the plant.

22. The method of any one of clauses 1 to 19 wherein the effectiveamount is an amount greater than about 1.0×10² CFU/ml or gram of thecomposition used to contact the plant to about 1.0×10³ CFU/ml or gram ofthe composition used to contact the plant.

23. The method of any one of clauses 1 to 22 wherein the Bacillus straincomprises live Bacillus cells or spores.

24. The method of clause 23 wherein the Bacillus strain comprises liveBacillus cells.

25. The method of clause 23 wherein the Bacillus strain comprisesspores.

26. The method of any one of clauses 1 to 25 wherein the composition isa liquid.

27. The method of any one of clauses 1 to 25 wherein the composition isdry.

28. The method of clause 27 wherein the composition is lyophilized.

29. The method of any one of clauses 1 to 28 wherein the contacting isby spraying the composition onto the plant.

30. The method of any one of clauses 1 to 28 wherein the contacting isby manually applying the composition onto the plant.

31. The method of any one of clauses 1 to 30 wherein the plant iscontacted by contacting the composition with a part of the plantselected from the group consisting of a leaf, a seed, a root, a flower,a shoot, a bud, and combinations thereof.

32. The method of any one of clauses 1 to 30 wherein the plant iscontacted by applying the composition to the soil from which the plantobtains nutrients.

33. The method of any one of clauses 1 to 32 wherein the plant is a palmplant.

34. A commercial package or a composition comprising an isolatedBacillus strain selected from the group consisting of Bacillus strainA12 (NRRL No. B-67516), a strain having all of the identifyingcharacteristics of Bacillus strain A12 (NRRL No. B-67516), Bacillusstrain V17 (NRRL No. B-67664), a strain having all of the identifyingcharacteristics of Bacillus strain V17 (NRRL No. B-67664), Bacillusstrain V18 (NRRL No. B-67665), a strain having all of the identifyingcharacteristics of Bacillus strain V18 (NRRL No. B-67665), Bacillusstrain 1607 (NRRL No. B-67666), a strain having all of the identifyingcharacteristics of Bacillus strain 1607 (NRRL No. B-67666), andcombinations thereof.

35. A commercial package or a composition comprising an isolatedBacillus strain used to inhibit a fungal disease or a bacterial diseasein a plant wherein the isolated Bacillus strain is selected from thegroup consisting of Bacillus strain 86 (NRRL No. B-50944), a strainhaving all of the identifying characteristics of Bacillus strain 86(NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), a strainhaving all of the identifying characteristics of Bacillus strain 300(NRRL No. B-50943), Bacillus strain 101 (NRRL No. B-67218), a strainhaving all of the identifying characteristics of Bacillus strain 101(NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), a strainhaving all of the identifying characteristics of Bacillus strain 235(NRRL No. B-67219), Bacillus strain 77 (NRRL No. B-67274), a strainhaving all of the identifying characteristics of Bacillus strain 77(NRRL No. B-67274), Bacillus strain 177 (NRRL No. B-67275), a strainhaving all of the identifying characteristics of Bacillus strain 177(NRRL No. B-67275), Bacillus strain 102 (NRRL No. B-67276), a strainhaving all of the identifying characteristics of Bacillus strain 102(NRRL No. B-67276), Bacillus strain ATC2 (NRRL No. B-67134), a strainhaving all of the identifying characteristics of Bacillus strain ATC2(NRRL No. B-67134), Bacillus strain A12 (NRRL No. B-67516), a strainhaving all of the identifying characteristics of Bacillus strain A12(NRRL No. B-67516), Bacillus strain V17 (NRRL No. B-67664), a strainhaving all of the identifying characteristics of Bacillus strain V17(NRRL No. B-67664), Bacillus strain V18 (NRRL No. B-67665), a strainhaving all of the identifying characteristics of Bacillus strain V18(NRRL No. B-67665), Bacillus strain 1607 (NRRL No. B-67666), a strainhaving all of the identifying characteristics of Bacillus strain 1607(NRRL No. B-67666), and combinations thereof.

36. The commercial package or composition of any one of clauses 34 to 35wherein the Bacillus strain is in the form of a concentrate.

37. The commercial package or composition of any one of clauses 34 to 35wherein the Bacillus strain is in the form of a superconcentrate.

38. The commercial package or composition of any one of clauses 34 to 37wherein the Bacillus strain is in a dry form.

39. The commercial package or composition of any one of clauses 34 to 37wherein the Bacillus strain is in a form selected from the groupconsisting of a powder and a liquid.

40. The commercial package or composition of any one of clauses 34 to 39further comprising a carrier for the Bacillus strain.

41. The commercial package or composition of clause 40 wherein thecarrier is selected from the group consisting of salt, a dextrin, andcombinations thereof.

42. The commercial package or composition of any one of clauses 34 to 41in a bag.

43. The commercial package or composition of clause 42 wherein the bagis a plastic bag.

44. The commercial package or composition of any one of clauses 34 to 43further comprising instructions for use of one or more of the Bacillusstrains.

45. The commercial package or composition of any one of clauses 42 to 43in a 20-pound bag.

46. The commercial package or composition of any one of clauses 42 to 43in a 50-pound bag.

47. The commercial package or composition of clause 39 wherein theBacillus strain is in powder form.

48. The commercial package or composition of clause 39 wherein theBacillus strain is in liquid form.

49. The commercial package or composition of any one of clauses 34 to 48wherein the Bacillus strain is in a container for commercial use.

50. The commercial package or composition of clause 49 wherein thecontainer comprises plastic.

51. The commercial package or composition of clause 49 wherein thecontainer comprises paper.

52. The commercial package or composition of any one of clauses 34 to 51further comprising a binder.

53. The commercial package or composition of clause 52 wherein thebinder is selected from the group consisting of clay, yeast cell wallcomponents, aluminum silicate, and glucan, or combinations thereof.

54. The commercial package or composition of any one of clauses 34 or 36to 53 wherein the Bacillus strain is used to inhibit a fungal disease ina plant.

55. The commercial package or composition of any one of clauses 34 or 36to 53 wherein the Bacillus strain is used to inhibit a bacterial diseasein a plant.

56. The method of clause 11 wherein the fungus is Cladosporiumcladosporioides.

57. The method of clause 11 wherein the fungus is Pichia manshurica.

58. The method of clause 11 wherein the fungus is Pichia kudriavzevii.

59. The method of clause 11 wherein the fungus is Aspergillus fumigatus.

60. The method of any one of clauses 1 to 33 wherein the bacterialdisease is caused by bacteria of the genus Ralstonia.

61. The method of clause 60 wherein the bacteria are Ralstoniasolanacearum.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the percent inhibition of growth of Ganoderma boninense,Ganoderma weberianum, and Fusarium oxysporum in broth culture bysupernatants from various Bacillus strains.

FIG. 2A shows that Bacillus strain 300 inhibited A. fumigatus biomassaccumulation following 24 hours of growth in a liquid broth assay(p=0.02). FIG. 2B. is a photograph following 24 hours of A. fumigatusgrowth in liquid broth (tube 1, left) or liquid broth with Bacillusstrain 300 supernatant (tube 2, right).

DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

Applicants have developed Bacillus strains, and combinations thereof,that can be used to treat plants to inhibit a fungal disease or abacterial disease. More particularly, the invention relates to isolatedBacillus strains, and strains having all of the identifyingcharacteristics of these strains, and combinations thereof, for a usecomprising the above-mentioned use. These fungal and bacterial diseasesdetrimentally affect plants and cause significant economic losses in thehorticulture, agricultural, and forestry industries, for example.

In one embodiment, a method of treating a plant to inhibit a fungaldisease or a bacterial disease is provided. The method comprisescontacting the plant with a composition comprising an effective amountof an isolated Bacillus strain selected from the group consisting ofBacillus strain 86 (NRRL No. B-50944), a strain having all of theidentifying characteristics of Bacillus strain 86 (NRRL No. B-50944),Bacillus strain 300 (NRRL No. B-50943), a strain having all of theidentifying characteristics of Bacillus strain 300 (NRRL No. B-50943),Bacillus strain 101 (NRRL No. B-67218), a strain having all of theidentifying characteristics of Bacillus strain 101 (NRRL No. B-67218),Bacillus strain 235 (NRRL No. B-67219), a strain having all of theidentifying characteristics of Bacillus strain 235 (NRRL No. B-67219),Bacillus strain 77 (NRRL No. B-67274), a strain having all of theidentifying characteristics of Bacillus strain 77 (NRRL No. B-67274),Bacillus strain 177 (NRRL No. B-67275), a strain having all of theidentifying characteristics of Bacillus strain 177 (NRRL No. B-67275),Bacillus strain 102 (NRRL No. B-67276), a strain having all of theidentifying characteristics of Bacillus strain 102 (NRRL No. B-67276),Bacillus strain ATC2 (NRRL No. B-67134), a strain having all of theidentifying characteristics of Bacillus strain ATC2 (NRRL No. B-67134),Bacillus strain A12 (NRRL No. B-67516), a strain having all of theidentifying characteristics of Bacillus strain A12 (NRRL No. B-67516),Bacillus strain V17 (NRRL No. B-67664), a strain having all of theidentifying characteristics of Bacillus strain V17 (NRRL No. B-67664),Bacillus strain V18 (NRRL No. B-67665), a strain having all of theidentifying characteristics of Bacillus strain V18 (NRRL No. B-67665),Bacillus strain 1607 (NRRL No. B-67666), a strain having all of theidentifying characteristics of Bacillus strain 1607 (NRRL No. B-67666),and combinations thereof, and inhibiting the fungal disease or thebacterial disease.

In another embodiment, a commercial package is provided. The commercialpackage comprises an isolated Bacillus strain selected from the groupconsisting of Bacillus strain A12 (NRRL No. B-67516), a strain havingall of the identifying characteristics of Bacillus strain A12 (NRRL No.B-67516), Bacillus strain V17 (NRRL No. B-67664), a strain having all ofthe identifying characteristics of Bacillus strain V17 (NRRL No.B-67664), Bacillus strain V18 (NRRL No. B-67665), a strain having all ofthe identifying characteristics of Bacillus strain V18 (NRRL No.B-67665), Bacillus strain 1607 (NRRL No. B-67666), a strain having allof the identifying characteristics of Bacillus strain 1607 (NRRL No.B-67666), and combinations thereof.

In yet another embodiment, a composition is provided comprising anisolated Bacillus strain selected from the group consisting of Bacillusstrain A12 (NRRL No. B-67516), a strain having all of the identifyingcharacteristics of Bacillus strain A12 (NRRL No. B-67516), Bacillusstrain V17 (NRRL No. B-67664), a strain having all of the identifyingcharacteristics of Bacillus strain V17 (NRRL No. B-67664), Bacillusstrain V18 (NRRL No. B-67665), a strain having all of the identifyingcharacteristics of Bacillus strain V18 (NRRL No. B-67665), Bacillusstrain 1607 (NRRL No. B-67666), a strain having all of the identifyingcharacteristics of Bacillus strain 1607 (NRRL No. B-67666), andcombinations thereof. In another aspect, this Bacillus straincomposition can be used to inhibit a fungal disease or a bacterialdisease in a plant.

In another embodiment, a commercial package or a composition is providedcomprising an isolated Bacillus strain used to inhibit a fungal diseaseor a bacterial disease in a plant wherein the isolated Bacillus strainis selected from the group consisting of Bacillus strain 86 (NRRL No.B-50944), a strain having all of the identifying characteristics ofBacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No.B-50943), a strain having all of the identifying characteristics ofBacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No.B-67218), a strain having all of the identifying characteristics ofBacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No.B-67219), a strain having all of the identifying characteristics ofBacillus strain 235 (NRRL No. B-67219), Bacillus strain 77 (NRRL No.B-67274), a strain having all of the identifying characteristics ofBacillus strain 77 (NRRL No. B-67274), Bacillus strain 177 (NRRL No.B-67275), a strain having all of the identifying characteristics ofBacillus strain 177 (NRRL No. B-67275), Bacillus strain 102 (NRRL No.B-67276), a strain having all of the identifying characteristics ofBacillus strain 102 (NRRL No. B-67276), Bacillus strain ATC2 (NRRL No.B-67134), a strain having all of the identifying characteristics ofBacillus strain ATC2 (NRRL No. B-67134), Bacillus strain A12 (NRRL No.B-67516), a strain having all of the identifying characteristics ofBacillus strain A12 (NRRL No. B-67516), Bacillus strain V17 (NRRL No.B-67664), a strain having all of the identifying characteristics ofBacillus strain V17 (NRRL No. B-67664), Bacillus strain V18 (NRRL No.B-67665), a strain having all of the identifying characteristics ofBacillus strain V18 (NRRL No. B-67665), Bacillus strain 1607 (NRRL No.B-67666), a strain having all of the identifying characteristics ofBacillus strain 1607 (NRRL No. B-67666), and combinations thereof.

The following clauses, and combinations thereof, provide variousadditional illustrative aspects of the invention described herein. Thevarious embodiments described in this section titled “DETAILEDDESCRIPTION OF ILLUSTRATIVE EMBODIMENTS” are applicable to any of thefollowing embodiments of the invention described in the numbered clausesbelow.

1. A method of treating a plant to inhibit a fungal disease or abacterial disease, the method comprising contacting the plant with acomposition comprising an effective amount of an isolated Bacillusstrain selected from the group consisting of Bacillus strain 86 (NRRLNo. B-50944), a strain having all of the identifying characteristics ofBacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No.B-50943), a strain having all of the identifying characteristics ofBacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No.B-67218), a strain having all of the identifying characteristics ofBacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No.B-67219), a strain having all of the identifying characteristics ofBacillus strain 235 (NRRL No. B-67219), Bacillus strain 77 (NRRL No.B-67274), a strain having all of the identifying characteristics ofBacillus strain 77 (NRRL No. B-67274), Bacillus strain 177 (NRRL No.B-67275), a strain having all of the identifying characteristics ofBacillus strain 177 (NRRL No. B-67275), Bacillus strain 102 (NRRL No.B-67276), a strain having all of the identifying characteristics ofBacillus strain 102 (NRRL No. B-67276), Bacillus strain ATC2 (NRRL No.B-67134), a strain having all of the identifying characteristics ofBacillus strain ATC2 (NRRL No. B-67134), Bacillus strain A12 (NRRL No.B-67516), a strain having all of the identifying characteristics ofBacillus strain A12 (NRRL No. B-67516), Bacillus strain V17 (NRRL No.B-67664), a strain having all of the identifying characteristics ofBacillus strain V17 (NRRL No. B-67664), Bacillus strain V18 (NRRL No.B-67665), a strain having all of the identifying characteristics ofBacillus strain V18 (NRRL No. B-67665), Bacillus strain 1607 (NRRL No.B-67666), a strain having all of the identifying characteristics ofBacillus strain 1607 (NRRL No. B-67666), and combinations thereof, andinhibiting the fungal disease or the bacterial disease.

2. The method of clause 1 wherein the Bacillus strain is selected fromthe group consisting of Bacillus strain 300 (NRRL No. B-50943), a strainhaving all of the identifying characteristics of Bacillus strain 300(NRRL No. B-50943), Bacillus strain 101 (NRRL No. B-67218), a strainhaving all of the identifying characteristics of Bacillus strain 101(NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), a strainhaving all of the identifying characteristics of Bacillus strain 235(NRRL No. B-67219), Bacillus strain 77 (NRRL No. B-67274), a strainhaving all of the identifying characteristics of Bacillus strain 77(NRRL No. B-67274), Bacillus strain A12 (NRRL No. B-67516), a strainhaving all of the identifying characteristics of Bacillus strain A12(NRRL No. B-67516), Bacillus strain 1607 (NRRL No. B-67666), a strainhaving all of the identifying characteristics of Bacillus strain 1607(NRRL No. B-67666), and combinations thereof.

3. The method of clause 1 or 2 wherein the Bacillus strain is selectedfrom the group consisting of Bacillus strain 300 (NRRL No. B-50943), astrain having all of the identifying characteristics of Bacillus strain300 (NRRL No. B-50943), Bacillus strain 235 (NRRL No. B-67219), a strainhaving all of the identifying characteristics of Bacillus strain 235(NRRL No. B-67219), Bacillus strain 1607 (NRRL No. B-67666), a strainhaving all of the identifying characteristics of Bacillus strain 1607(NRRL No. B-67666), and combinations thereof.

4. The method of clause 2 wherein the Bacillus strain is Bacillus strain300 (NRRL No. B-50943).

5. The method of clause 2 where the Bacillus strain is Bacillus strain235 (NRRL No. B-67219).

6. The method of clause 2 wherein the Bacillus strain is Bacillus strain1607 (NRRL No. B-67666).

7. The method of clause 2 wherein the Bacillus strain is Bacillus strain101 (NRRL No. B-67218).

8. The method of clause 2 wherein the Bacillus strain is Bacillus strain77 (NRRL No. B-67274).

9. The method of any one of clauses 1 to 8 wherein the Bacillus strainhas antifungal activity.

10. The method of any one of clauses 1 to 9 wherein the fungal diseaseis caused by a fungus of a genus selected from the group consisting ofGanoderma, Phytophthora, Fusarium, and combinations thereof.

11. The method of clause 10 wherein the fungal disease is caused by afungus selected from the group consisting of Ganoderma boninense,Ganoderma mirabile, Ganoderma weberianum, Phytophthora palmivora,Fusarium kyushuense, Fusarium oxysporum, Fusarium nelsonii, Cladosporiumcladosporioides, Pichia manshurica, Pichia kudriavzevii, Aspergillusfumigatus, and combinations thereof.

12. The method of clause 11 wherein the fungus is Ganoderma boninense.

13. The method of clause 11 wherein the fungus is Ganoderma mirabile.

14. The method of clause 11 wherein the fungus is Ganoderma weberianum.

15. The method of clause 11 wherein the fungus is Phytophthorapalmivora.

16. The method of clause 11 wherein the fungus is Fusarium kyushuense.

17. The method of clause 11 wherein the fungus is Fusarium oxysporum.

18. The method of clause 11 wherein the fungus is Fusarium nelsonii.

19. The method of any one of clauses 1 to 18 further comprising treatingthe plant with a different Bacillus strain, a lactic acid bacterialstrain, and combinations thereof.

20. The method of any one of clauses 1 to 19 wherein the effectiveamount of the Bacillus strain is about 1.0×10² CFU/ml or gram of thecomposition used to contact the plant to about 1.0×10⁸ CFU/ml or gram ofthe composition used to contact the plant.

21. The method of any one of clauses 1 to 19 wherein the effectiveamount of the Bacillus strain is about 1.0×10² CFU/ml or gram of thecomposition used to contact the plant to about 1.0×10⁴ CFU/ml or gram ofthe composition used to contact the plant.

22. The method of any one of clauses 1 to 19 wherein the effectiveamount is an amount greater than about 1.0×10² CFU/ml or gram of thecomposition used to contact the plant to about 1.0×10³ CFU/ml or gram ofthe composition used to contact the plant.

23. The method of any one of clauses 1 to 22 wherein the Bacillus straincomprises live Bacillus cells or spores.

24. The method of clause 23 wherein the Bacillus strain comprises liveBacillus cells.

25. The method of clause 23 wherein the Bacillus strain comprisesspores.

26. The method of any one of clauses 1 to 25 wherein the composition isa liquid.

27. The method of any one of clauses 1 to 25 wherein the composition isdry.

28. The method of clause 27 wherein the composition is lyophilized.

29. The method of any one of clauses 1 to 28 wherein the contacting isby spraying the composition onto the plant.

30. The method of any one of clauses 1 to 28 wherein the contacting isby manually applying the composition onto the plant.

31. The method of any one of clauses 1 to 30 wherein the plant iscontacted by contacting the composition with a part of the plantselected from the group consisting of a leaf, a seed, a root, a flower,a shoot, a bud, and combinations thereof.

32. The method of any one of clauses 1 to 30 wherein the plant iscontacted by applying the composition to the soil from which the plantobtains nutrients.

33. The method of any one of clauses 1 to 32 wherein the plant is a palmplant.

34. A commercial package or a composition comprising an isolatedBacillus strain selected from the group consisting of Bacillus strainA12 (NRRL No. B-67516), a strain having all of the identifyingcharacteristics of Bacillus strain A12 (NRRL No. B-67516), Bacillusstrain V17 (NRRL No. B-67664), a strain having all of the identifyingcharacteristics of Bacillus strain V17 (NRRL No. B-67664), Bacillusstrain V18 (NRRL No. B-67665), a strain having all of the identifyingcharacteristics of Bacillus strain V18 (NRRL No. B-67665), Bacillusstrain 1607 (NRRL No. B-67666), a strain having all of the identifyingcharacteristics of Bacillus strain 1607 (NRRL No. B-67666), andcombinations thereof.

35. A commercial package or a composition comprising an isolatedBacillus strain used to inhibit a fungal disease or a bacterial diseasein a plant wherein the isolated Bacillus strain is selected from thegroup consisting of Bacillus strain 86 (NRRL No. B-50944), a strainhaving all of the identifying characteristics of Bacillus strain 86(NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), a strainhaving all of the identifying characteristics of Bacillus strain 300(NRRL No. B-50943), Bacillus strain 101 (NRRL No. B-67218), a strainhaving all of the identifying characteristics of Bacillus strain 101(NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), a strainhaving all of the identifying characteristics of Bacillus strain 235(NRRL No. B-67219), Bacillus strain 77 (NRRL No. B-67274), a strainhaving all of the identifying characteristics of Bacillus strain 77(NRRL No. B-67274), Bacillus strain 177 (NRRL No. B-67275), a strainhaving all of the identifying characteristics of Bacillus strain 177(NRRL No. B-67275), Bacillus strain 102 (NRRL No. B-67276), a strainhaving all of the identifying characteristics of Bacillus strain 102(NRRL No. B-67276), Bacillus strain ATC2 (NRRL No. B-67134), a strainhaving all of the identifying characteristics of Bacillus strain ATC2(NRRL No. B-67134), Bacillus strain A12 (NRRL No. B-67516), a strainhaving all of the identifying characteristics of Bacillus strain A12(NRRL No. B-67516), Bacillus strain V17 (NRRL No. B-67664), a strainhaving all of the identifying characteristics of Bacillus strain V17(NRRL No. B-67664), Bacillus strain V18 (NRRL No. B-67665), a strainhaving all of the identifying characteristics of Bacillus strain V18(NRRL No. B-67665), Bacillus strain 1607 (NRRL No. B-67666), a strainhaving all of the identifying characteristics of Bacillus strain 1607(NRRL No. B-67666), and combinations thereof.

36. The commercial package or composition of any one of clauses 34 to 35wherein the Bacillus strain is in the form of a concentrate.

37. The commercial package or composition of any one of clauses 34 to 35wherein the Bacillus strain is in the form of a superconcentrate.

38. The commercial package or composition of any one of clauses 34 to 37wherein the Bacillus strain is in a dry form.

39. The commercial package or composition of any one of clauses 34 to 37wherein the Bacillus strain is in a form selected from the groupconsisting of a powder and a liquid.

40. The commercial package or composition of any one of clauses 34 to 39further comprising a carrier for the Bacillus strain.

41. The commercial package or composition of clause 40 wherein thecarrier is selected from the group consisting of salt, a dextrin, andcombinations thereof.

42. The commercial package or composition of any one of clauses 34 to 41in a bag.

43. The commercial package or composition of clause 42 wherein the bagis a plastic bag.

44. The commercial package or composition of any one of clauses 34 to 43further comprising instructions for use of one or more of the Bacillusstrains.

45. The commercial package or composition of any one of clauses 42 to 43in a 20-pound bag.

46. The commercial package or composition of any one of clauses 42 to 43in a 50-pound bag.

47. The commercial package or composition of clause 39 wherein theBacillus strain is in powder form.

48. The commercial package or composition of clause 39 wherein theBacillus strain is in liquid form.

49. The commercial package or composition of any one of clauses 34 to 48wherein the Bacillus strain is in a container for commercial use.

50. The commercial package or composition of clause 49 wherein thecontainer comprises plastic.

51. The commercial package or composition of clause 49 wherein thecontainer comprises paper.

52. The commercial package or composition of any one of clauses 34 to 51further comprising a binder.

53. The commercial package or composition of clause 52 wherein thebinder is selected from the group consisting of clay, yeast cell wallcomponents, aluminum silicate, and glucan, or combinations thereof.

54. The commercial package or composition of any one of clauses 34 or 36to 53 wherein the Bacillus strain is used to inhibit a fungal disease ina plant.

55. The commercial package or composition of any one of clauses 34 or 36to 53 wherein the Bacillus strain is used to inhibit a bacterial diseasein a plant.

56. The method of clause 11 wherein the fungus is Cladosporiumcladosporioides.

57. The method of clause 11 wherein the fungus is Pichia manshurica.

58. The method of clause 11 wherein the fungus is Pichia kudriavzevii.

59. The method of clause 11 wherein the fungus is Aspergillus fumigatus.

60. The method of any one of clauses 1 to 33 wherein the bacterialdisease is caused by bacteria of the genus Ralstonia.

61. The method of clause 60 wherein the bacteria are Ralstoniasolanacearum.

In various embodiments, the Bacillus strain for use in accordance withthe methods, commercial packages, and compositions described herein canbe selected from the group consisting of Bacillus strain 86 (NRRL No.B-50944), a strain having all of the identifying characteristics ofBacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No.B-50943), a strain having all of the identifying characteristics ofBacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No.B-67218), a strain having all of the identifying characteristics ofBacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No.B-67219), a strain having all of the identifying characteristics ofBacillus strain 235 (NRRL No. B-67219), Bacillus strain 77 (NRRL No.B-67274), a strain having all of the identifying characteristics ofBacillus strain 77 (NRRL No. B-67274), Bacillus strain 177 (NRRL No.B-67275), a strain having all of the identifying characteristics ofBacillus strain 177 (NRRL No. B-67275), Bacillus strain 102 (NRRL No.B-67276), a strain having all of the identifying characteristics ofBacillus strain 102 (NRRL No. B-67276), Bacillus strain ATC2 (NRRL No.B-67134), a strain having all of the identifying characteristics ofBacillus strain ATC2 (NRRL No. B-67134), Bacillus strain A12 (NRRL No.B-67516), a strain having all of the identifying characteristics ofBacillus strain A12 (NRRL No. B-67516), Bacillus strain V17 (NRRL No.B-67664), a strain having all of the identifying characteristics ofBacillus strain V17 (NRRL No. B-67664), Bacillus strain V18 (NRRL No.B-67665), a strain having all of the identifying characteristics ofBacillus strain V18 (NRRL No. B-67665), Bacillus strain 1607 (NRRL No.B-67666), a strain having all of the identifying characteristics ofBacillus strain 1607 (NRRL No. B-67666), and combinations thereof.

Bacillus strain MDG 101 and Bacillus strain MDG 235 were deposited onJan. 4, 2016 at the Agricultural Research Service Culture Collection(NRRL), National Center for Agricultural Utilization Research,Agricultural Research Service, USDA, 1815 North University Street,Peoria, Ill. 61604-3999, and were given accession numbers B-67218 andB-67219, respectively. Bacillus strain MGL77, Bacillus strain MGL177,and Bacillus strain MGL102 were deposited on June 7, 2016 at theAgricultural Research Service Culture Collection (NRRL), National Centerfor Agricultural Utilization Research, Agricultural Research Service,USDA, 1815 North University Street, Peoria, Ill. 61604-3999, and weregiven accession numbers B-67274, B-67275, and B-67276, respectively. Thedeposits were made under the provisions of the Budapest Treaty on theInternational Recognition of the Deposit of Microorganisms for thePurposes of Patent Procedure. The NRRL strain designations are MDG 101,MDG 235, MGL77, MGL177, and MGL102 which are equivalent to Bacillusstrain 101, 235, 77, 177, and 102 respectively, as referred to in theapplication. Bacillus strain MDG86 and Bacillus strain MDG300 weredeposited on Mar. 14, 2014 at the Agricultural Research Service CultureCollection (NRRL), National Center for Agricultural UtilizationResearch, Agricultural Research Service, USDA, 1815 North UniversityStreet, Peoria, Ill. 61604-3999, and were given accession numbersB-50944 and B-50943, respectively. The deposits were made under theprovisions of the Budapest Treaty on the International Recognition ofthe Deposit of Microorganisms for the Purposes of Patent Procedure. TheNRRL strain designations are MDG86 and MDG300, which are equivalent toBacillus strain 86 and 300, respectively, as referred to in theapplication.

Bacillus strain ATC2 was deposited on Sep. 16, 2015 at the AgriculturalResearch Service Culture Collection (NRRL), National Center forAgricultural Utilization Research, Agricultural Research Service, USDA,1815 North University Street, Peoria, Ill. 61604-3999, and was givenaccession number B-67134. The deposit was made under the provisions ofthe Budapest Treaty on the International Recognition of the Deposit ofMicroorganisms for the Purposes of Patent Procedure. The NRRL straindesignation is ATC2, which is equivalent to Bacillus strain ATC2, asreferred to in the application.

Bacillus strain MDGA12 was deposited on Sep. 14, 2017 at theAgricultural Research Service Culture Collection (NRRL), InternationalDepository Authority, 1815 North University Street, Peoria, Ill. 61604,and was given accession number B-67516. The deposit was made under theprovisions of the Budapest Treaty on the International Recognition ofthe Deposit of Microorganisms for the Purposes of Patent Procedure. TheNRRL strain designation is MDGA12, which is equivalent to Bacillusstrain A12, as referred to in the application.

Bacillus strains V17, V18, and 1607 were deposited on Aug. 14, 2018 atthe Agricultural Research Service Culture Collection (NRRL),International Depository Authority, 1815 North University Street,Peoria, Ill. 61604, and were given accession numbers B-67664, B-67665,and B-67666, respectively. The deposits were made under the provisionsof the Budapest Treaty on the International Recognition of the Depositof Microorganisms for the Purposes of Patent Procedure. The NRRL straindesignations are MDGV17, MDGV18, and MDG1607, which are equivalent toBacillus strain V17, V18, and 1607, respectively, as referred to in theapplication. Bacillus strain V17 is a Bacillus pumilus strain andBacillus strain 1607 is a Bacillus amyloliquefaciens strain, strains 102and 177 are Bacillus pumilus strains, strain 77 is a Bacilluslicheniformis strain, and all other strains described in the applicationare Bacillus subtilus strains.

In one illustrative aspect, any of these strains can be used to treatplants to inhibit fungal or bacterial infections or diseases alone or incombination in the form of a liquid or dry spray to be applied to theplant or to the soil from which the plant obtains nutrients, or in theform of a liquid or dry composition for manual application to the plantor to the soil from which the plant obtains nutrients. In oneembodiment, multiple strains are used to treat the plants in combinationin a single composition. In another embodiment, multiple strains areused to treat the plant in combination in separate compositions.

As used herein “a strain having all of the identifying characteristicsof” Bacillus strain 86, 300, 101, 235, 77, 177, 102, A12, ATC2, V17,V18, or 1607 can be a mutant strain having all of the identifyingcharacteristics of Bacillus strain 86, 300, 101, 235, 77, 177, 102, A12,ATC2, V17, V18, or 1607 (e.g., a DNA fingerprint based on DNA analysisthat corresponds to the DNA fingerprint of Bacillus strain 86, 300, 101,235, 77, 177, 102, A12, ATC2, V17, V18, or 1607, enzyme activities thatcorrespond to Bacillus strain 86, 300, 101, 235, 77, 177, 102, A12,ATC2, V17, V18, or 1607, antimicrobial activity that corresponds toBacillus strain 86, 300, 101, 235, 77, 177, 102, A12, ATC2, V17, V18, or1607, antibiotic sensitivity and tolerance profiles that correspond toBacillus strain 86, 300, 101, 235, 77, 177, 102, A12, ATC2, V17, V18, or1607, or combinations thereof). In alternate embodiments, the mutationcan be a natural mutation, or a genetically engineered mutation. Inanother embodiment, “a strain having all of the identifyingcharacteristics of” Bacillus strain 86, 300, 101, 235, 77, 177, 102,A12, ATC2, V17, V18, or 1607 can be a strain, for example, produced byisolating one or more plasmids from Bacillus strain 86, 300, 101, 235,77, 177, 102, A12, ATC2, V17, V18, or 1607 and introducing the one ormore plasmids into another bacterium, such as another Bacillus strain,as long as the one or more plasmids contain DNA that provides theidentifying characteristics of Bacillus strain 86, 300, 101, 235, 77,177, 102, A12, ATC2, V17, V18, or 1607 (e.g., a DNA fingerprint based onDNA analysis that corresponds to the DNA fingerprint of Bacillus strain86, 300, 101, 235, 77, 177, 102, A12, ATC2, V17, V18, or 1607).

In another embodiment, one or more of the Bacillus strains described inthe preceding paragraphs (e.g., Bacillus strain 86, 300, 101, 235, 77,177, 102, A12, ATC2, V17, V18, or 1607) can be used to treat plantsalong with another bacterial strain selected from the group consistingof a different Bacillus strain, a lactic acid bacterial strain, andcombinations thereof. In still another embodiment, the additionalBacillus strain can be selected from the group consisting of Bacillusamyloliquefaciens, Bacillus licheniformis, Bacillus pumilus, otherBacillus strains, and combinations thereof. In yet another embodiment,one or more of the Bacillus strains described in the precedingparagraphs (e.g., Bacillus strain 86, 300, 101, 235, 77, 177, 102, A12,ATC2, V17, V18, or 1607) can be used to treat plants along with anyother bacterial strain effective to treat plants to inhibit a fungal orbacterial infection or a fungal or bacterial disease.

As used herein the terms “inhibit”, “inhibiting”, and “inhibited” meanto reduce or eliminate the symptoms of a fungal or bacterial disease ora fungal or bacterial infection for the plant or a human, or to reduceor eliminate the number, or an activity, of a fungus or bacterium of atype that causes a fungal or bacterial disease or a fungal or bacterialinfection, and which is found in association with the plant or human.Exemplary fungal diseases that can be inhibited using the methods,commercial packages, and compositions described herein are basal stemrot disease, bud rot, wilt, soft rot, Cladosporium rot, andaspergillosis. An exemplary bacterial disease is bacterial wilt causedby Ralstonia solanacearum. The Bacillus strains described herein canalso prevent off-odors on fruit, and wine spoilage resulting in loss ofwine quality and financial loss.

The compositions described herein can be used to treat plants for anyperiod of time that is effective to inhibit a fungal or bacterialdisease or a fungal or bacterial infection and/or to control thedetrimental effects of a fungal or bacterial disease or a fungal orbacterial infection. For example, in one embodiment treatment of theplants can occur daily, bi-weekly, three times a week, four times aweek, five times a week, once weekly, monthly, or for any suitable timeor using any suitable protocol for inhibiting a fungal or bacterialdisease or a fungal or bacterial infection. The time periods fortreatment of the plants are non-limiting and it should be appreciatedthat any time period or treatment schedule determined to be effective toinhibit a fungal or bacterial disease or a fungal or bacterial infectionand/or control the detrimental effects of a fungal or bacterial diseaseor a fungal or bacterial infection may be used.

In various illustrative embodiments, the Bacillus strain (e.g., Bacillusstrain 86, 300, 101, 235, 77, 177, 102, A12, ATC2, V17, V18, or 1607),or any other bacterial strains added in addition to Bacillus strain 86,300, 101, 235, 77, 177, 102, A12, ATC2, V17, V18, or 1607, can be usedto treat plants at about 1.0×10² CFU, about 1.0×10² CFU to about 1.0×10³CFU, about 1.0×10² CFU to about 1.0×10⁴ CFU, about 1.0×10² CFU to about1.0×10⁵ CFU, about 1.0×10² CFU to about 1.0×10⁶ CFU, about 1.0×10² CFUto about 1.0×10⁷ CFU. about 1.0×10² CFU to about 1.0×10⁸ CFU, about1.0×10² CFU to about 1.0×10⁹ CFU, about 1.0×10² CFU to about 1.0×10¹⁰CFU, about 1.0×10³ CFU to about 5.0×10¹² CFU or at about 1.0×10³ CFU toabout 1.0×10¹⁰ CFU. In other embodiments, the Bacillus strain (e.g.,Bacillus strain 86, 300, 101, 235, 77, 177, 102, A12, ATC2, V17, V18, or1607) can be used to treat the plants at an amount greater than or about1.0×10² CFU, at greater than or about 1.0×10³ CFU, at greater than orabout 1.1×10³ CFU, at greater than or about 1.25×10³ CFU, at greaterthan or about 1.5×10³ CFU, at greater than or about 1.75×10³ CFU, atgreater than or about 1.0×10⁴ CFU, at greater than or about 2.0×10⁴ CFU,at greater than or about 3.0×10⁴ CFU, at greater than or about 4.0×10⁴CFU, at greater than or about 5.0×10⁴ CFU, at greater than or about6.0×10⁴ CFU, at greater than or about 7.0×10⁴ CFU, at greater than orabout 8.0×10⁴ CFU, at greater than or about 1.0×10⁵ CFU, at greater thanor about 1.0×10⁶ CFU, at greater than or about 1.0×10⁷ CFU, at greaterthan or about 1.0×10⁸ CFU, at greater than or about 1.0×10⁹ CFU, atgreater than or about 1.0×10¹⁰ CFU, at greater than or about 1.0×10¹¹CFU, or at greater than or about 1.0×10¹² CFU. In all of the embodimentsusing “CFU”, the CFU can be CFU/gram or CFU/ml of the liquid or powderedformulation applied to the plants.

In various illustrative aspects, the fungi that can be inhibited includea fungus of a genus selected from the group consisting of Ganoderma,Phytophthora, Fusarium, Cladosporium, Pichia, and Aspergillus, andcombinations thereof, or a fungus selected from the group consisting ofGanoderma boninense, Ganoderma mirabile, Ganoderma weberianum,Phytophthora palmivora, Fusarium kyushuense, Fusarium oxysporum,Fusarium nelsonii, Cladosporium cladosporioides, Pichia manshurica,Pichia kudriavzevii, Aspergillus fumigatus, and combinations thereof. Insome embodiments described herein, the Bacillus strains (e.g., Bacillusstrain 86, 300, 101, 235, 77, 177, 102, A12, ATC2, V17, V18, or 1607)can have antifungal activity. Such antifungal activity can be against,for example, a fungus of a genus selected from the group consisting ofGanoderma, Phytophthora, Fusarium, Cladosporium, Pichia, andAspergillus, and combinations thereof, or a fungus selected from thegroup consisting of Ganoderma boninense, Ganoderma mirabile, Ganodermaweberianum, Phytophthora palmivora, Fusarium kyushuense, Fusariumoxysporum, Fusarium nelsonii, Cladosporium cladosporioides, Pichiamanshurica, Pichia kudriavzevii, Aspergillus fumigatus, and combinationsthereof. As used herein, Pichia are yeast, but can be considered a typeof fungus. Other types of yeast (i.e., considered fungi) that may beinhibited by the Bacillus species described herein are Saccharomycesspecies, Kluyveromyces species, Torulaspora species, Schizosaccharomycesspecies, Hansenula species, Torulopsis species, Candida species, andKarwinskia species. In yet another embodiment, any fungal disease of aplant can be inhibited by the Bacillus strains 86, 300, 101, 235, 77,177, 102, A12, ATC2, V17, V18, or 1607, and a strain having all of theidentifying characteristics of the Bacillus strains, or combinationsthereof, described herein. In still another embodiment, any bacterialdisease of a plant can be inhibited by the Bacillus strains 86, 300,101, 235, 77, 177, 102, A12, ATC2, V17, V18, or 1607, and a strainhaving all of the identifying characteristics of the Bacillus strains,or combinations thereof, described herein, such as bacterial wilt causedby Ralstonia solanacearum.

In one embodiment, a commercial package is provided comprising anisolated Bacillus strain selected from the group consisting of Bacillusstrain 86, 300, 101, 235, 77, 177, 102, A12, ATC2, V17, V18, or 1607, astrain having all of the identifying characteristics of any of theseBacillus strains, and combinations thereof.

In another embodiment, a composition is provided comprising an isolatedBacillus strain selected from the group consisting of Bacillus strain86, 300, 101, 235, 77, 177, 102, A12, ATC2, V17, V18, or 1607, a strainhaving all of the identifying characteristics of any of these Bacillusstrains, and combinations thereof.

In yet another embodiment, a commercial package or a composition isprovided comprising an isolated Bacillus strain used to inhibit a fungalor bacterial disease in a plant wherein the isolated Bacillus strain isselected from the group consisting of Bacillus strain 86 (NRRL No.B-50944), a strain having all of the identifying characteristics ofBacillus strain 86 (NRRL No. B-50944), Bacillus strain 300 (NRRL No.B-50943), a strain having all of the identifying characteristics ofBacillus strain 300 (NRRL No. B-50943), Bacillus strain 101 (NRRL No.B-67218), a strain having all of the identifying characteristics ofBacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No.B-67219), a strain having all of the identifying characteristics ofBacillus strain 235 (NRRL No. B-67219), Bacillus strain 77 (NRRL No.B-67274), a strain having all of the identifying characteristics ofBacillus strain 77 (NRRL No. B-67274), Bacillus strain 177 (NRRL No.B-67275), a strain having all of the identifying characteristics ofBacillus strain 177 (NRRL No. B-67275), Bacillus strain 102 (NRRL No.B-67276), a strain having all of the identifying characteristics ofBacillus strain 102 (NRRL No. B-67276), Bacillus strain ATC2 (NRRL No.B-67134), a strain having all of the identifying characteristics ofBacillus strain ATC2 (NRRL No. B-67134), Bacillus strain A12 (NRRL No.B-67516), a strain having all of the identifying characteristics ofBacillus strain A12 (NRRL No. B-67516), Bacillus strain V17 (NRRL No.B-67664), a strain having all of the identifying characteristics ofBacillus strain V17 (NRRL No. B-67664), Bacillus strain V18 (NRRL No.B-67665), a strain having all of the identifying characteristics ofBacillus strain V18 (NRRL No. B-67665), Bacillus strain 1607 (NRRL No.B-67666), a strain having all of the identifying characteristics ofBacillus strain 1607 (NRRL No. B-67666), and combinations thereof.

In these embodiments the Bacillus strain can be in the form of, forexample, a liquid or a dry composition, such as a suspension or alyophilized composition or a powder, a freeze-dried composition, a gel,or a pellet, for example. In one illustrative embodiment, the Bacillusstrain can be in the form of a powder or a liquid, and can be formulatedto be sprayed on or manually applied to the plant or to the soil fromwhich the plant obtains nutrients. In other embodiments, the Bacillusstrain can be mixed with fertilizer, mixed with soil, or used as a seedcoating for contacting the plant or a plant part. Any suitable methodknown in the art for contacting the plant or the soil with the Bacillusstrain can be used to achieve any of the effective amounts of Bacillusstrain 86, 300, 101, 235, 77, 177, 102, A12, ATC2, V17, V18, or 1607, ora strain having all of the identifying characteristics of any of theseBacillus strains, or combinations thereof, described herein. In oneaspect, the Bacillus strain can comprise live Bacillus cells or spores.In another illustrative embodiment, the plant can be contacted with theBacillus strain by contacting the Bacillus strain with a part of theplant selected from the group consisting of a leaf, a seed, a root, aflower, a shoot, a bud, and combinations thereof.

In illustrative aspects, Bacillus strain 86, 300, 101, 235, 77, 177,102, A12, ATC2, V17, V18, or 1607, or a strain having all of theidentifying characteristics of any of these Bacillus strains, andcombinations thereof, can be in the form of a commercial package, or anyother suitable composition. In another illustrative embodiment, theBacillus strain(s) in the commercial package, or other composition canbe in the form of a concentrate (e.g., about 1×10⁸ to about 5×10⁹ CFU/g)or a superconcentrate (e.g., about 1×10¹⁰ to about 5×10¹² CFU/g). Inanother embodiment, the Bacillus strain(s) in the commercial package orother composition can be in a dry form (e.g., a powder), a liquid form,a freeze-dried form, or in the form of a gel, or any other suitable formfor application to plants or the soil from which the plant obtainsnutrients.

In another illustrative embodiment, the Bacillus strain(s) in thecommercial package or composition can further comprise a carrier for theBacillus strain(s). In various embodiments, the carrier can be selectedfrom the group consisting of bran, rice hulls, a salt, mineral oil, adextrin (e.g., maltodextrin), whey, sugar, limestone, dried starch,sodium silico aluminate, vegetable oil, and combinations thereof. Inanother embodiment, the carrier can be any suitable carrier known in theart for a composition for application to plants to treat plants having afungal or bacterial disease or infection. In another embodiment, theBacillus strain(s) in the commercial package or composition can furthercomprise a binder such as clay, yeast cell wall components, aluminumsilicate, glucan, or other known binders, and/or micronutrients,including but not limited to, nitrogen and phosphorus. Any of thesecomponents can be exogenously added (i.e., not naturally present incombination with the Bacillus strains) and any of the compositionsdescribed herein can be present in nutrient compositions for plants.

In yet other embodiments, the commercial package or compositioncomprising an isolated Bacillus strain can be for inhibiting a fungal ora bacterial disease, and the Bacillus strain can be selected from thegroup consisting of Bacillus strain 86 (NRRL No. B-50944), a strainhaving all of the identifying characteristics of Bacillus strain 86(NRRL No. B-50944), Bacillus strain 300 (NRRL No. B-50943), a strainhaving all of the identifying characteristics of Bacillus strain 300(NRRL No. B-50943), Bacillus strain 101 (NRRL No. B-67218), a strainhaving all of the identifying characteristics of Bacillus strain 101(NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), a strainhaving all of the identifying characteristics of Bacillus strain 235(NRRL No. B-67219), Bacillus strain 77 (NRRL No. B-67274), a strainhaving all of the identifying characteristics of Bacillus strain 77(NRRL No. B-67274), Bacillus strain 177 (NRRL No. B-67275), a strainhaving all of the identifying characteristics of Bacillus strain 177(NRRL No. B-67275), Bacillus strain 102 (NRRL No. B-67276), a strainhaving all of the identifying characteristics of Bacillus strain 102(NRRL No. B-67276), Bacillus strain ATC2 (NRRL No. B-67134), a strainhaving all of the identifying characteristics of Bacillus strain ATC2(NRRL No. B-67134), Bacillus strain A12 (NRRL No. B-67516), a strainhaving all of the identifying characteristics of Bacillus strain A12(NRRL No. B-67516), Bacillus strain V17 (NRRL No. B-67664), a strainhaving all of the identifying characteristics of Bacillus strain V17(NRRL No. B-67664), Bacillus strain V18 (NRRL No. B-67665), a strainhaving all of the identifying characteristics of Bacillus strain V18(NRRL No. B-67665), Bacillus strain 1607 (NRRL No. B-67666), a strainhaving all of the identifying characteristics of Bacillus strain 1607(NRRL No. B-67666), and combinations thereof.

In another embodiment, the commercial package or composition can be in acontainer for commercial use. In various embodiments the container canbe, for example, a bag (e.g., a 20-pound bag, a 50-pound bag, a 2-ouncebag, a 1-pound bag, or a 1-kilogram bag), a pouch, a drum, a bottle, ora box. In illustrative aspects, the container comprising the Bacillusstrain(s) can comprise plastic, metal, foil, paper, fiber, or cardboard(e.g., a plastic pail, a paper bag, a foil bag, a fiber drum, etc.). Thecommercial package can further comprise instructions for use of one ormore of the Bacillus strains.

The following examples are for illustrative purposes only. The examplesare non-limiting, and are not intended to limit the invention in anyway.

EXAMPLE 1 Culture Conditions

All fungal cultures were obtained from ATCC and include Ganodermaboninense ATCC 204074, Ganoderma weberianum ATCC 76753, Ganodermamirabile ATCC 76757, 76537, Phytophthora palmivora ATCC 26008,Phytophthora palmivora ATCC 52161, Fusarium oxysporum ATCC 36870,Fusarium kyushuense ATCC 56750, and Fusarium nelsonii 201410. Ganodermaspecies were cultured on yeast mold agar, Phytophthora species werecultured on tomato juice agar, and Fusarium species were cultured onpotato dextrose agar. Bacillus strains 86, 300, 101, 235, 77, 177, 102,A12, ATC2, V17, V18, or 1607 were cultured on tryptic soy agar.

EXAMPLE 2 Antifungal Plate Assay

Plates were inoculated with an agar plug of fungal growth placed in thecenter of the plate. Bacillus culture was struck on the plate in twoparallel lines on either side of the agar plug at a distance of 3 cm.The plates were incubated at 30° C. until the fungal culture exceeded 6cm in diameter. Data was recorded at 6 days for all strains except F.kyushuense, which was recorded at 12 days. Fungal growth was measured,and inhibition scores were assigned for each Bacillus strain. Scores of−, +, ++, or +++ were defined as no inhibition, inhibition of 1 cmfungal growth, inhibition of 2 cm, and inhibition of 3 cm, respectively.

Several Bacillus strains had strong antifungal activity against all sixfungal strains (see Table 1 below). The top six strains had at leastmoderate activity against all fungal strains, and were further evaluatedfor antifungal activity in a broth assay.

TABLE 1 Inhibition of fungal growth by Bacillus strains in an antifungalplate assay. Inhibition of more than 3 cm of fungal growth is indicatedby “+++”, 2 to 3 cm inhibition indicated by “++”, 1 to 2 cm inhibitionindicated by “+”, and no inhibition is indicated by “−.” Gano- Gano-Gano- Phyto- Phyto- derma derma derma phthora phthora Fusarium FusariumFusarium boninense mirabile weberianum palmivora 1 palmivora 2kyushuense oxysporum nelsonii 77 +++ + +++ ++ +++ ++ ++ ++ 1607 ++ + ++++++ +++ ++ ++ ++ ATC2 +++ + +++ ++ +++ ++ ++ ++ 300 +++ ++ +++ ++ +++ ++++ ++ 101 +++ + +++ ++ +++ ++ ++ ++ 235 +++ ++ +++ ++ +++ ++ ++ + 86++ + + ++ +++ + ++ + V18 ++ + − ++ +++ + ++ + A12 ++ + + ++ +++ + + +V17 ++ + ++ + +++ − + + 102 − + − − +++ − − − 177 − + − − ++ − − −

EXAMPLE 3 Antifungal Broth Assay

The top performing Bacillus strains in the plate assay were evaluatedusing a broth method. An agar plug of fungal growth was used toinoculate 20 ml of broth media containing 10% cell-free Bacillussupernatant. Supernatants were prepared by growing Bacillus in trypticsoy broth for 24 hours, centrifuging the culture for 20 minutes at 4000rpm, and filter-sterilizing the supernatant with a 0.22 μm filter.Fungal cultures were grown at 30° C. for two weeks. Fungal growth wasfiltered and allowed to dry completely. Total dry fungal weight wasrecorded, and inhibition was reported as a percentage of the reductionin growth weight compared to controls. G. mirabile and P. palmivorastrains were excluded from the broth assay as they did not grow well inbroth media.

F. nelsonii and F. kyushuense were not inhibited at all by any Bacillusspecies (data not shown). However, all six Bacillus strains inhibited G.boninense by over 40%, the pathogen responsible for causing basal stemrot in oil palm plants (see FIG. 1 —strain “AlgT2” in FIG. 1 is the sameas strain ATC2). Bacillus strain 235, 300, and 1607 also had strongactivity against G. weberianum. Bacillus strains 101 and 300 had thebest activity against F. oxysporum.

Example 4 Isolation And Identification of Additional Pathogenic orSpoilage Fungi

Additional fungal isolates were obtained from molding or rottingvegetation including spoiled corn and hay silage, and were isolated bydilution plating on acidified potato dextrose agar incubated at roomtemperature (19-21° C.), protected from light, for 2-5 days untildistinct isolate growth was observed. DNA was extracted from fungalisolates using the Nucleospin Microbial DNA extraction kit(Machery-Nagel, Düren, Germany), employing a bead-beating step for celllysis. The fungal ribosomal intergenic spacer (ITS) region was amplifiedusing primers ITS1 (TCCGTAGGTGAACCTGCGG)(SEQ ID NO: 1) and ITS4(TCCTCCGCTTATTGATATGC)(SEQ ID NO: 2)(White, et al., Amplification anddirect sequencing of fungal ribosomal RNA genes for phylogenetics, pp.315-322, In: PCR Protocols: A Guide to Methods and Applications, eds.Innis, et al., Academic Press, Inc., New York (1990), incorporatedherein by reference for the amplification method described in thisexample), following recommended amplification parameters. Product sizewas determined by gel electrophoresis, the PCR product was cleaned upwith the Qiagen MinElute PCR purification kit (Hilden, Germany), and DNAsequencing of each purified PCR product was performed by EurofinsGenomics (Louisville, Ky.). Sequences were trimmed for quality, inChromas 2.6 (Technelysium, South Brisbane, Australia) and identified tothe closest species match using a nucleotide BLAST search (NationalCenter for Biotechnology Information, Bethesda, Md.).

EXAMPLE 5 Antifungal Cross-Streak Assay Using Plant and Spoilage Fungi

Strain-specific antifungal activities of Bacillus strains were testedusing agar cross-streak antimicrobial susceptibility methods known inthe art and described below. Briefly, Bacillus strains 77, 86, 300, 101,235, and V18 were inoculated from frozen glycerol stocks in a single 1cm wide vertical streak down the center of a non-acidified potatodextrose agar plate. Plates were incubated aerobically for 24 hours at37° C., or until a confluent streak of Bacillus growth was present.Isolates of Cladosporium, Pichia and A. fumigatus were struckhorizontally from the center of the plate, beginning the streak within 1mm of the Bacillus growth and streaking out to the plate perimeter.Plates were incubated at 25° C. under aerobic conditions for 3 days forPichia yeasts and 5 days for the remaining fungal isolates. Zones ofinhibition were measured where fungal isolate growth was inhibited bythe Bacillus strain. Table 2 shows antimicrobial screening utilizing theagar cross streak method against isolates of Cladosporiumcladosporioides, Pichia manshurica, Pichia kudriavzevii and Aspergillusfumigatus. Scores of −, +, ++, or +++ were defined as no inhibition,inhibition of 2-5 mm fungal growth, inhibition of 5-10 mm, andinhibition of >10 mm, respectively.

Several Bacillus strains had broad antifungal activity against allfungal strains tested (see Table 2 below). The top performing Bacillusstrain was further evaluated for antifungal activity in a broth assay.

TABLE 2 Inhibition of fungal growth by Bacillus strains in an antifungalcross-streak assay. Inhibition of more than 10 mm of fungal growth isindicated by “+++”, 5 to 10 mm inhibition indicated by “+”, 2 to 5 mminhibition indicated by “+”, and less than 2 mm inhibition is indicatedby “−.” Fungal isolate Pichia Pichia Pichia Pichia Pichia Pichia Clado-Asper- kudria- kudria- mans- mans- mans- mans- sporium gillus vzeviivzevii churica churica churica churica clado- fumi- Isolate 1 Isolate 2Isolate I Isolate 2 Isolate 3 Isolate 4 sporioides gatus Bacillus 77 ++++ ++ ++ ++ ++ +++ + strain 86 − − − − − − NA NA 101 + + + + + ++ ++ −235 ++ ++ +++ ++ ++ ++ +++ + 300 ++ ++ ++ ++ ++ ++ +++ ++ V18 − − − − −− +++ −

EXAMPLE 6 Aspergillus Fumigatus Antifungal Broth Assay

The top performing Bacillus strain in the plate assay (Bacillus strain300) was evaluated using a broth method to determine efficacy ofinhibition of A. fumigatus in a liquid matrix. The supernatant wasprepared by growing Bacillus strain 300 in tryptic soy broth for 24hours at 32° C., centrifuging the culture for 20 minutes at 4000 rpm,and filter-sterilizing the supernatant with a 0.22 μm filter.Aspergillus fumigatus was propagated in potato dextrose broth to producea suspension of approximately 10⁵ spores/ml. A 0.1 ml amount of thisinoculum was used to inoculate 5 ml of potato dextrose broth mixed with1 ml of sterile-filtered supernatant from Bacillus strain 300 or with 1ml of sterile culture broth (Control). Nine replicates of each treatmentwere prepared and all replicates were incubated for 24 hr at 37° C.Replicates were pooled into triplicate samples, fungal biomass wassedimented via centrifugation at 4000 rpm for 20 minutes, washed withsterile water to remove media residue, dried on sterile filters andweighed to determine fungal biomass. The photograph shown in FIG. 2B wastaken at the end of the 24 hour growth period and the tubes are fromleft to right, control and Bacillus strain 300 treatment. Total dryfungal weight was recorded, and inhibition was reported as a percentageof the reduction in growth weight compared to controls. Bacillus strain300 significantly inhibited A. fumigatus biomass (p=0.02), the speciesassociated with acceleration of soft rot in many plant species and whichis responsible for opportunistic infections in humans (see FIG. 2A).

EXAMPLE 7 Inhibition of Ralstonia Solanacearum by Bacillus Strains

Ralstonia solanacearum is a soilborne pathogen that causes bacterialwilt in a variety of crops. Strains of Ralstonia were isolated from soiland tested for inhibition of the isolated Ralstonia strains by Bacillusspecies. Soil samples were plated for Ralstonia species onsemi-selective media. Modified SMSA medium contained 1 g/L casein, 10g/L peptone, 5 ml/L glycerol, and 17 g/L agar at pH 7.0 (Elphinstone etal., 1996). After autoclaving, media was supplemented with 5 mg/Lcrystal violet, 100 mg/L polymyxin B sulfate, 25 mg/L bacitracin, 5 mg/Lchloromycetin, 0.5 mg/L penicillin, and 100 mg/L cycloheximide.Presumptive Ralstonia isolates were identified as Ralstonia species by16S sequencing. Bacillus species were tested for antimicrobial activityagainst the Ralstonia species using a cross streak method. The zone ofinhibition between the Bacillus and Ralstonia streak was measured in mm.

As shown in Table 3 below, all Bacillus species tested showed someinhibition of all three Ralstonia isolates. Bacillus species have thepotential to be used as biocontrol agents against Ralstonia species.

TABLE 3 Inhibition (mm) of Ralstonia species by Bacillus. The zone ofinhibition between the Bacillus and Ralstonia streak was measured in mm.Inhibition of more than 10 mm growth is indicated by “+++”, 6 to 9 mminhibition is indicated by “++”, 1 to 5 mm inhibition is indicated by“+”, and no inhibition is indicated by “−”. Bacillus Ralstonia RalstoniaRalstonia strain species 1 species 2 species 3 300 +++ +++ +++ 235 +++++ +++ 1607 ++ +++ +++ AlgT2 ++ +++ +++ (equivalent to ATC2) 77 ++ +++++ V18 + ++ ++ 101 ++ ++ ++ V17 ++ ++ +

What is claimed is:
 1. A method of treating a plant to inhibit a fungaldisease or a bacterial disease, the method comprising preparing acomposition comprising one or more isolated Bacillus strains wherein thecomposition is prepared to form a powder, a gel, a pellet, a liquid, ora freeze-dried composition, and contacting the plant with thecomposition wherein the one or more isolated Bacillus strains comprisean effective amount of isolated Bacillus strain 300 (NRRL No. B-50943),wherein Bacillus strain 300 (NRRL No. B-50943) inhibits the fungaldisease or the bacterial disease.
 2. The method of claim 1 wherein theplant is further treated with a Bacillus strain selected from the groupconsisting of Bacillus strain 101 (NRRL No. B-67218), Bacillus strain235 (NRRL No. B-67219), Bacillus strain 77 (NRRL No. B-67274), Bacillusstrain A12 (NRRL No. B-67516), Bacillus strain 1607 (NRRL No. B-67666),and combinations thereof, wherein the additional Bacillus strains alsoinhibit the fungal disease or the bacterial disease.
 3. The method ofclaim 1 wherein the plant is further treated with a Bacillus strainselected from the group consisting of Bacillus strain 235 (NRRL No.B-67219), Bacillus strain 1607 (NRRL No. B-67666), and combinationsthereof, wherein the additional Bacillus strains also inhibit the fungaldisease or the bacterial disease.
 4. The method of claim 2 wherein theBacillus strain is Bacillus strain 1607 (NRRL No. B-67666).
 5. Themethod of claim 1 wherein the Bacillus strain has antifungal activity.6. The method of claim 5 wherein the fungal disease is caused by afungus of a genus selected from the group consisting of Ganoderma,Phytophthora, Fusarium, and combinations thereof.
 7. The method of claim5 wherein the fungal disease is caused by a fungus selected from thegroup consisting of Ganoderma boninense, Ganoderma mirabile, Ganodermaweberianum, Phytophthora palmivora, Fusarium kyushuense, Fusariumoxysporum, Fusarium nelsonii, Cladosporium cladosporioides, Pichiamanshurica, Pichia kudriavzevii, Aspergillus fumigatus, and combinationsthereof.
 8. The method of claim 7 wherein the fungus is Ganodermaboninense.
 9. The method of claim 7 wherein the fungus is Ganodermamirabile.
 10. The method of claim 7 wherein the fungus is Ganodermaweberianum.
 11. The method of claim 7 wherein the fungus is Phytophthorapalmivora.
 12. The method of claim 7 wherein the fungus is Fusariumkyushuense.
 13. The method of claim 7 wherein the fungus is Fusariumoxysporum.
 14. The method of claim 7 wherein the fungus is Fusariumnelsonii.
 15. The method of claim 7 wherein the fungus is Cladosporiumcladosporioides.
 16. The method of claim 7 wherein the fungus is Pichiamanshurica.
 17. The method of claim 7 wherein the fungus is Pichiakudriavzevii.
 18. The method of claim 7 wherein the fungus isAspergillus fumigatus.
 19. The method of claim 1 wherein the bacterialdisease is caused by bacteria of the genus Ralstonia.
 20. The method ofclaim 19 wherein the bacteria are Ralstonia solanacearum.
 21. The methodof claim 1 wherein the plant is further treated with a Bacillus strainselected from the group consisting of Bacillus strain 101 (NRRL No.B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 77(NRRL No. B-67274), Bacillus strain 177 (NRRL No. B-67275), Bacillusstrain 102 (NRRL No. B-67276), Bacillus strain ATC2 (NRRL No. B-67134),Bacillus strain A12 (NRRL No. B-67516), Bacillus strain V17 (NRRL No.B-67664), Bacillus strain V18 (NRRL No. B-67665), Bacillus strain 1607(NRRL No. B-67666), and combinations thereof, wherein the additionalBacillus strains also inhibit the fungal disease or the bacterialdisease.
 22. A method of treating a plant to inhibit a fungal disease ora bacterial disease, the method comprising preparing a compositioncomprising one or more isolated Bacillus strains wherein the compositionis prepared to form a powder, a gel, a pellet, a liquid, or afreeze-dried composition, and contacting the plant with the compositionwherein the one or more isolated Bacillus strains comprise an effectiveamount of isolated Bacillus strain 1607 (NRRL No. B-67666), whereinBacillus strain 1607 (NRRL No. B-67666) inhibits the fungal disease orthe bacterial disease.
 23. The method of claim 22 wherein the plant isfurther treated with a Bacillus strain selected from the groupconsisting of Bacillus strain 300 (NRRL No. B-50943), Bacillus strain101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillusstrain 77 (NRRL No. B-67274), Bacillus strain Al2 (NRRL No. B-67516),and combinations thereof, wherein the additional Bacillus strains alsoinhibit the fungal disease or the bacterial disease.
 24. The method ofclaim 22 wherein the plant is further treated with a Bacillus strainselected from the group consisting of Bacillus strain 300 (NRRL No.B-50943), Bacillus strain 235 (NRRL No. B-67219), and combinationsthereof, wherein the additional Bacillus strains also inhibit the fungaldisease or the bacterial disease.
 25. The method of claim 23 wherein theBacillus strain is Bacillus strain 300 (NRRL No. B-50943).
 26. Themethod of claim 22 wherein the Bacillus strain has antifungal activity.27. The method of claim 22 wherein the fungal disease is caused by afungus of a genus selected from the group consisting of Ganoderma,Phytophthora, Fusarium, and combinations thereof.
 28. The method ofclaim 22 wherein the fungal disease is caused by a fungus selected fromthe group consisting of Ganoderma boninense, Ganoderma mirabile,Ganoderma weberianum, Phytophthora palmivora, Fusarium kyushuense,Fusarium oxysporum, Fusarium nelsonii, Cladosporium cladosporioides,Pichia manshurica, Pichia kudriavzevii, Aspergillus fumigatus, andcombinations thereof.
 29. The method of claim 28 wherein the fungus isFusarium oxysporum.
 30. A method of treating a plant to inhibit a fungaldisease or a bacterial disease, the method comprising preparing acomposition comprising one or more isolated Bacillus strains wherein thecomposition is prepared to form a powder, a gel, a pellet, a liquid, ora freeze-dried composition, and contacting the plant with thecomposition wherein the one or more isolated Bacillus strains comprisean effective amount of an isolated Bacillus strain selected from thegroup consisting of Bacillus strain 86 (NRRL No. B-50944), Bacillusstrain 300 (NRRL No. B-50943), Bacillus strain ATC2 (NRRL No. B-67134),Bacillus strain A12 (NRRL No. B-67516), Bacillus strain V17 (NRRL No.B-67664), Bacillus strain V18 (NRRL No. B-67665), Bacillus strain 1607(NRRL No. B-67666), and combinations thereof, and inhibiting the fungaldisease or the bacterial disease.
 31. The method of claim 30 wherein theBacillus strain has antifungal activity.
 32. The method of claim 30wherein the fungal disease is caused by a fungus of a genus selectedfrom the group consisting of Ganoderma, Phytophthora, Fusarium, andcombinations thereof.
 33. The method of claim 30 wherein the fungaldisease is caused by a fungus selected from the group consisting ofGanoderma boninense, Ganoderma mirabile, Ganoderma weberianum,Phytophthora palmivora, Fusarium kyushuense, Fusarium oxysporum,Fusarium nelsonii, Cladosporium cladosporioides, Pichia manshurica,Pichia kudriavzevii, Aspergillus fumigatus, and combinations thereof.34. The method of claim 33 wherein the fungus is Fusarium oxysporum.